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在酵母中引入额外氧化还原辅因子以实现 F 依赖性生物转化。

Equipping with an Additional Redox Cofactor Allows F-Dependent Bioconversions in Yeast.

机构信息

Molecular Enzymology Group, University of Groningen, Nijenborgh 4, 9747AG Groningen, The Netherlands.

出版信息

ACS Synth Biol. 2024 Mar 15;13(3):921-929. doi: 10.1021/acssynbio.3c00718. Epub 2024 Feb 12.

Abstract

Industrial application of the natural deazaflavin cofactor F has high potential for the enzymatic synthesis of high value compounds. It can offer an additional range of chemistry to the use of well-explored redox cofactors such as FAD and their respective enzymes. Its limited access through organisms that are rather difficult to grow has urged research on the heterologous production of F using more industrially relevant microorganisms such as . In this study, we demonstrate the possibility of producing this cofactor in a robust and widely used industrial organism, , by the heterologous expression of the F pathway. Through careful selection of involved enzymes and some optimization, we achieved an F yield of ∼1.3 μmol/L, which is comparable to the yield of natural F producers. Furthermore, we showed the potential use of F-producing for F-dependent bioconversions by carrying out the whole-cell conversion of tetracycline. As the first demonstration of F synthesis and use for bioconversion in a eukaryotic organism, this study contributes to the development of versatile bioconversion platforms.

摘要

天然去氮黄素辅因子 F 在工业上的应用在高价值化合物的酶促合成方面具有很大的潜力。它可以为使用经过充分探索的氧化还原辅因子(如 FAD 及其各自的酶)提供更多的化学选择。由于其来源的生物体生长困难,因此迫切需要研究使用更具工业相关性的微生物(如 )来异源生产 F。在这项研究中,我们通过 F 途径的异源表达,证明了在一种强大且广泛使用的工业生物体 中生产这种辅因子的可能性。通过仔细选择相关酶并进行一些优化,我们实现了约 1.3 μmol/L 的 F 产量,与天然 F 产生菌的产量相当。此外,我们还通过全细胞转化四环素展示了生产 F 的 用于 F 依赖性生物转化的潜力。作为在真核生物中进行 F 合成和用于生物转化的首次展示,这项研究为多功能生物转化平台的发展做出了贡献。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df46/10949242/bbb03dfd39e9/sb3c00718_0001.jpg

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